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<p>Lymphocystis disease (LCD), induced by the lymphocystis disease virus (LCDV), results in losses across more than 140 fish species. In this study, we present a rapid, convenient, and highly specific detection assay based on recombinase polymerase amplification–lateral flow dipstick (RPA–LFD) technology to identify LCDV. This assay targets the highly conserved major capsid protein (<i>MCP</i>) of LCDV using specific and sensitive primers and probes. The optimized protocol necessitates merely 15&#xa0;min at 43&#xa0;°C, achieving a detection threshold of a minimum of 13&#xa0;copies/μL. The entire RPA–LFD assay is over 1 h more rapid than the quantitative real-time polymerase chain reaction (qPCR) method, while achieving a high level of sensitivity that is close to that of qPCR and demonstrating no cross-reactivity with other aquatic viruses. The isothermal amplification RPA–LFD technique provides a straightforward, rapid, and reliable approach that can potentially enhance field diagnosis of LCDV, particularly in resource-constrained settings.</p>

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Establishment of the recombinase polymerase amplification–lateral flow dipstick (RPA–LFD) detection technique for rapid detection of lymphocystis disease virus (LCDV)

  • 期刊论文

  • 发表日期：

    2025年9月12日

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  • Kaijie Yu,
  • Yitong Zhang,
  • Ziyang He,
  • Yufeng Liu,
  • Zhongwei He,
  • Wei Cao,
  • Yuqin Ren,
  • Guixing Wang,
  • Yufen Wang,
  • Jilun Hou

摘要

Lymphocystis disease (LCD), induced by the lymphocystis disease virus (LCDV), results in losses across more than 140 fish species. In this study, we present a rapid, convenient, and highly specific detection assay based on recombinase polymerase amplification–lateral flow dipstick (RPA–LFD) technology to identify LCDV. This assay targets the highly conserved major capsid protein (MCP) of LCDV using specific and sensitive primers and probes. The optimized protocol necessitates merely 15 min at 43 °C, achieving a detection threshold of a minimum of 13 copies/μL. The entire RPA–LFD assay is over 1 h more rapid than the quantitative real-time polymerase chain reaction (qPCR) method, while achieving a high level of sensitivity that is close to that of qPCR and demonstrating no cross-reactivity with other aquatic viruses. The isothermal amplification RPA–LFD technique provides a straightforward, rapid, and reliable approach that can potentially enhance field diagnosis of LCDV, particularly in resource-constrained settings.